Rodent Cremaster Preparation Device for Intravital Microscopy

Abstract

Provided herein is a device useful for preparing a cremaster muscle of a specimen for intravital microscopy.

1 . A device useful for securing a cremaster muscle of a specimen during intravital microscopy comprising:
 a) a surgery tray,   b) immobilizers, and   c) a temperature-controlled cremaster muscle mount, wherein the mount comprises a temperature sensor, a heating element, a viewing plate, a well formed by a gasket for physiological buffer, a plurality of stand-offs, and a plurality of hooks and sutures,   
       wherein the mount is positioned onto the surgery tray to hold the cremaster muscle of the specimen and wherein the hooks are positioned inside the well and are suitable for spreading the cremaster muscle, whereby the cremaster muscle is viewable under a microscope. 
     
     
         2 . The device of  claim 1  further comprising a micropump and tubing, wherein the micropump and tubing circulate physiological buffer over the cremaster muscle within the well. 
     
     
         3 . The device of  claim 1  further comprising a heating pad. 
     
     
         4 . The device of  claim 1 , wherein the specimen is a mammal. 
     
     
         5 . The device of  claim 4 , wherein the mammal is a rodent. 
     
     
         6 . The device of  claim 1 , wherein the surgery tray is fabricated from a material selected from the group consisting of a plastic, a polymer, a glass, a metal or a combination thereof. 
     
     
         7 . The device of  claim 1 , wherein the surgery tray comprises a magnetic element for variable positioning and fixing of the temperature-controlled cremaster muscle mount to the tray. 
     
     
         8 . The device of  claim 1 , wherein the immobilizers comprise pegs and bands, wherein the pegs are useful for immobilizing the ilium and rear legs and wherein the bands are useful for immobilizing the head and body. 
     
     
         9 . The device of  claim 1 , wherein the well for physiological buffer surrounds a region of the viewing plate to immerse the cremaster muscle in buffer. 
     
     
         10 . The device of  claim 1 , wherein the hooks are fabricated from wire. 
     
     
         11 . A mounting device useful for securing a cremaster muscle of a specimen during intravital microscopy comprising:
 a) a plate,   b) a plate mount,   c) a plurality of stand-offs,   d) a temperature sensor,   e) a heating element,   f) a gasket, wherein the gasket creates a well, and   g) a plurality of hooks and sutures.   
     
     
         12 . The device of  claim 11 , wherein the plate is glass. 
     
     
         13 . The device of  claim 11 , wherein the plate mount is a material selected from the group consisting of plastic, metal, or glass. 
     
     
         14 . The device of  claim 11 , wherein the stand-off is a material selected from the group consisting of plastic, metal, or glass. 
     
     
         15 . The device of  claim 11 , wherein the stand-off is adjustable in height. 
     
     
         16 . The device of  claim 11 , wherein the stand-off comprises a means for attachment to a tray. 
     
     
         17 . The device of  claim 16 , wherein the means for attachment to the tray is magnetic. 
     
     
         18 . The device of  claim 11 , wherein the gasket is a material selected from the group consisting of a rubber, silicone, plastic, metal, or glass. 
     
     
         19 . The device of  claim 11 , wherein the hooks are fabricated from wire. 
     
     
         20 . The device of  claim 11 , further comprising a micropump and tubing, wherein the micropump and tubing circulate physiological buffer over the cremaster muscle within the well. 
     
     
         21 . A method of using the device of  claim 11 , wherein a cremaster muscle of a specimen is positioned on the plate within the gasket and is stretched to a thin layer using the hooks. 
     
     
         22 . A method of preparing a cremaster muscle for intravital microscopy comprising the steps of:
 a) providing a specimen,   b) placing the specimen on a surgery tray,   c) immobilizing the specimen using immobilizers situated in specific positions on the   tray,   d) isolating the specimen's cremaster muscle from the scrotum,   e) incising and positioning the cremaster muscle onto a temperature-controlled cremaster muscle mount,   f) inserting hooks attached to sutures positioned in a circular array into the cremaster, and   g) spreading the cremaster muscle using the hooks and sutures to a thin layer.